A técnica de inseminação artificial (IA) por laparoscopia em ovinos foi utilizada pela primeira vez em 1982, o que tornou viável a utilização econômica do sêmen congelado nesta espécie. Os experimentos foram realizados com o objetivo de determinar as taxas de prenhez de ovelhas inseminadas por laparoscopia, utilizando 50 x 106 espermatozóides descongelados e capacitados in vitro. No experimento 1, foi utilizado sêmen de um reprodutor e 67 ovelhas com estros sincronizados, divididas aleatoriamente entre os grupos: sêmen congelado (SC1) e sêmen congelado capacitado in vitro (SCC1). As ovelhas foram submetidas à IA, com 50x106 espermatozóides, 56 e 60 horas, respectivamente, após a retirada das esponjas impregnadas com progesterona. No grupo SCC1, a taxa de prenhez foi de 48,39% (15/31) significativamente superior a do grupo SC1 de 25% (9/36). No experimento 2, utilizou-se um reprodutor e 100 ovelhas com os estros sincronizados, distribuídas aleatoriamente entre três grupos: sêmen fresco (SF2), SC2 e SCC2. Nos grupos SF2 e SC2 as fêmeas foram inseminadas com 100x106 espermatozóides e 56 horas após a retirada da progesterona. No grupo das fêmeas inseminadas com espermatozóides capacitados in vitro, utilizou-se 50x106 espermatozóides e um intervalo de 60 horas após retirada da progesterona. As taxas de prenhez diagnosticadas nos diferentes grupos experimentais foram: SF2 60,9% (25/41), SC2 56,70% (17/30) e SCC2 44,90% (13/29), não existindo diferença estatística entre os grupos. Os experimentos realizados mostraram a viabilidade doemprego, na espécie ovina, da IA por laparoscopia utilizando 50x106 espermatozóides descongelados e capacitados in vitro. / The ovine artificial insemination (AI) by laparoscopy was described the first time in 1982, since this time frozen semen became a tool for use under field conditions. Today one research goal is to achieve high preganancies rates after AI using a reduced number of frozen/thawed spermatozoa per dosis. The experiments were carried out with the objective to determine the pregnancy rate of ewes inseminated by laparoscopy, using 50 x 106 thawed and in vitro capacited spermatozoa. In the experiment 1 was used semen of one fertily ram and 67 ewes with synchronized estrus. The females were randomly distributed among two experimental groups: inseminated with frozen semen (SC1) or frozen and in vitro capacitated semen (SCC1). The ewes were submitted to the AI with 50x106 spermatozoa, 56 (SC1) or 60 (SCC1) hours, respectively, after retreat of the intra vaginal device impregnated with progesterona. In the group SCC1 the pregnancy rate was 48,39% (15/31) significantly different from the 25% (9/36) observed in the group SC1. In the experiment 2 was used one fertily ram and 100 ewes with synchronized estrus, distributed randomly among three groups: inseminated with fresh semen (SF2), 100 x 106 spermatozoa, SC2, 100 x 106 frozen spermatozoa and SCC2, 50 x 106 frozen and in vitro capacited spermatozoa. The moment of the laparoscopic AI was the same as used in the experiment 1: 56 (SF2 and SC2) or 60 (SCC2) hours, respectively, after retreat of the intra vaginal device impregnated with progesterona. The observed pregnancy rates were similar among the experimental groups: SF2 60,9% (25/41), SC2 56,70% (17/30) and SCC2 44,90% (13/29). Our experiments showed the ability of frozen/thawed and in vitro capacitated ovine spermatozoa to in vivo fertilize and promote the development of pregancies. In conclusion, itis possible to use 50x106 frozen/thawed and in vitro capacited spermatozoa for ovine laparoscopic AI.

Submitted by (edna.saturno@ufrpe.br) on 2016-07-29T11:50:01Z No. of bitstreams: 1 Allan Vieira dos Santos Marques.pdf: 717930 bytes, checksum: aa978139a85c2cfa9f5a7df5d8379ad4 (MD5) / Made available in DSpace on 2016-07-29T11:50:01Z (GMT). No. of bitstreams: 1 Allan Vieira dos Santos Marques.pdf: 717930 bytes, checksum: aa978139a85c2cfa9f5a7df5d8379ad4 (MD5) Previous issue date: 2010-02-26 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / The aim of this research was to know serum and liver concentrations of Cu, Mo, Fe and Zn in sheep and goats slaughtered in the semiarid region of the state of Pernambuco, Northeastern Brazil, during the rainy and dry seasons, and to determine if Cu deficiency that occurs in the region is primary or secondary to high levels of Mo and/or Fe. Serum and liver samples from 141 goats and 141 sheep were submitted to wet digestion in nitric-percloric acid and analyzed by coupled plasma atomic absorption spectrophotometry. The mean concentration of serum Cu in sheep was 9.22±2.34 μmol/L in the dry period and 10.03±2.89 μmol/L in the rainy period, but females had significantly higher values (10.92±2.56 μmol/L) than males (9.42±2.67 μmol/L). Mean Cu concentrations in the liver were 185.1±88.28 mg per kg of dry matter (mg/kg) in the dry period and 137.95±87.04 mg/kg in the rainy period, and females had significantly higher concentrations (218.61±93.49 mg/kg) than males (133.45±92.37 mg/kg). In goats serum concentrations of Cu were of 11.74±2.36 μmol/L in the dry period and 11.05±2.74 μmol/L in the rainy period, and females (12.10±2.67 μmol/L) had higher concentrations than males (10.14±1.80 μmol/L). Liver concentrations were 155.2±93.40 mg/kg in the dry period and of 149.37±62.33 mg/kg in the rainy period. Mean serum concentrations of Cu in sheep (9.85±2.71 μmol/L) were significantlyt lower than in goats (11.37±2.57 μmol/L), but there were no differences in liver concentrations of Cu between sheep (158.45±83.05 mg/kg) and goats (152.46±79.58 mg/kg). Based in Cu serum and liver concentrations, and in the sporadic occurrence of enzootic ataxia, Cu suplementation is recommended in grazing sheep and goats during the dry and the rainy seasons. In sheep, serum concentrations of Fe were 36.99±15 μmol/L in the dry season and 34.52±14.86 μmol/L in the rainy season, with significant differences between females (29.92±11.06 μmol/L) and males (37.85±15.68 μmol/L). Liver Fe concentrations were significantly lower in the dry season (186.20±30.02 mg/kg) than in the rainy season (229.25±83.15 mg/kg), with no differences between females and males. In goats, serum concentrations of Fe were 25.61±9.35 μmol/L in the dry season and 24.57±6.96 μmol/L in the rainy season, and liver concentrations were of 170.52±56.91 mg/kg in the dry season and 139.73±51.30 mg/kg in the rainy season; there were no significant differences between females and males. Serum levels of Fe were significantly higher in sheep (35.58±14.89 μmol/L) than in goats (25.06±8.10 μmol/L). Also Fe concentrations of the liver were higher in sheep (210.53±121.99 mg/kg) than in goats (156.1±55.99 mg/kg). Considering that serum and hepatic concentrations of Fe are between normal ranges or marginal it is suggested that the low Cu concentrations are not related with high Fe ingestion. In sheep serum concentrations of Mo were 0.33±0.18 μmol/L in the dry season and 0.3±0.15 μmol/L in the rainy season and liver concentrations were 8.27±3.97 in the dry season and 7.96±4.07 in the rainy season. In goats, serum concentrations of Mo were 0.31±0.12 μmol/L in the dry season and 0.26±0.10 μmol/L in the rainy season, and liver concentrations were 7.17±4.09 mg/kg in the dry season and 5.8±4.14 mg/kg in the rainy season. There were no significant differences in serum and liver Mo concentrations between sheep and goats and between males and females. Despite the variations found in this study and also in reference values, it is suggested that the low or marginal Cu concentrations are not due to high Mo intake. In sheep serum concentrations of Zn were significantly lower during the dry season (9.79±3.02 μmol/L) than in the rainy season (13.48±7.22 μmol/L). Also Zn concentrations in the liver were significantly lower in the dry season (109.96±30.23 mg/kg) than in rainy season (139.1±60.45 mg/kg). In goats, serum Zn concentrations were 12.01±9.61 μmol/L in the dry season and 11.6±4.93 μmol/L in the rainy season, and liver concentrations were 134.2±26.58 mg/kg in the dry season and 131.45±76.55 mg/kg in the rainy season. There were no significant differences between serum and liver concentrations of Zn between sheep and goats or between males and females. Taking into account that serum and liver Zn concentrations in sheep and goats are considered marginal, supplementation with this mineral it suggested in grazing sheep and goats, mainly during the dry season. / Objetivou-se conhecer os teores de Cu, Mo, Fe e Zn em soro e fígado de ovinos e caprinos, nas épocas de chuva e seca, criadas no semi-árido do estado de Pernambuco, e estabelecer se a carência de Cu é causada por deficiência primária de Cu ou secundária à ingestão de quantidades excessivas de Fe ou Mo. Amostras de soro e fígado de 141 ovinos e 141 caprinos foram submetidas à digestão úmida em ácido nítrico-perclórico e analisados em espectrofotômetro de absorção atômica acoplado a plasma. A concentração sérica média de Cu em ovinos foi de 9,22±2,34 μmol/L na seca e 10,03±2,89 μmol/L na chuva, sendo que as fêmeas tiveram valores significativamente maiores (10,92±2,56 μmol/L) do que os machos (9,42±2,67 μmol/L). Os valores de Cu em fígado foram de 185,1±88,28 mg/kg da MS na seca e 137,95±87,04 mg/kg na chuva e as fêmeas tiveram valores significativamente maiores (218,61±93,49 mg/kg) do que os machos (133,45±92,37 mg/kg). Em caprinos os valores séricos de Cu foram de 11,74±2,36 μmol/L na seca e 11,05±2,74 μmol/L na chuva, sendo que as fêmeas tiveram maiores valores (12,10±2,67 μmol/L) que os machos (10,14±1,80 μmol/L). Os teores do fígado foram de 155,2±93,40 mg/kg na seca e 149,37±62,33 mg/kg na chuva. Os teores séricos de cobre em ovinos foram significativamente menores que o dos caprinos, mas não se observaram diferenças nos teores hepáticos de Cu entre ovinos e caprinos. Baseado nesses valores, considerados marginais, e na ocorrência de surtos de ataxia enzoótica em caprinos e ovinos na região, recomenda-se a suplementação com Cu, em animais a campo, tanto na seca quanto na chuva. Os teores séricos de Fe em ovinos foram de 36,99±15 μmol/L na seca e de 34,52±14,86 μmol/L na chuva, diferindo entre fêmeas (29,92±11,06 μmol/L) e machos (37,85±15,68 μmol/L). Os teores do fígado foram significativamente menores na seca (186,20±30,02 mg/kg) do que na chuva (229,25±83,15 mg/kg), não havendo diferenças entre machos e fêmeas. Em caprinos os teores séricos de Fe foram de 25,61±9,35 μmol/L na seca e de 24,57±6,96 μmol/L na chuva e os teores do fígado de 170,52±56,91 mg/kg na seca e de 139,73±51,30 mg/kg na chuva. Nesta espécie não houve diferenças nos teores séricos e hepáticos de Fe entre machos e fêmeas. Os teores séricos e hepáticos de Fe foram significativamente maiores em ovinos do que em caprinos. Considerando que as concentrações séricas e hepáticas de Fe encontram-se dentro da normalidade ou são marginais sugere-se que as concentrações marginais de Cu não estejam diretamente relacionadas com o excesso de Fe. Os teores séricos de Mo em ovinos foram de 0,33±0,18 μmol/L na seca e 0,3±0,15 μmol/L na chuva e os teores no fígado de 8,27±3,97 na seca e 7,96±4,07 na chuva. Os teores séricos de Mo em caprinos foram de 0,31±0,12 μmol/L na seca e de 0,26±0,10 μmol/L na chuva e os do fígado de 7,17±4,09 mg/kg na seca e 5,8±4,14 mg/kg na chuva. Os teores médios de Mo de soro e fígado em ovinos, 0,31±0,17 μmol/L e 8,10±4,01 mg/kg, respectivamente, foram semelhantes aos teores séricos e hepáticos de caprinos, de 0,28±0,11 μmol/L e 6,53±4,13 mg/kg, respectivamente. Tanto em ovinos quanto em caprinos não houve diferenças significantes nos teores do soro e fígado entre machos e fêmeas. Apesar das variações encontradas tanto neste trabalho quanto nos dados da literatura os resultados sugerem que os teores marginais de Cu não são devidos a teores altos de Mo. Em ovinos os valores de Zn no soro foram significativamente diferentes na seca (9,79±3,02 μmol/L) do que na chuva (13,48±7,22 μmol/L). No fígado os valores de Zn foram, também, significativamente diferentes na seca (109,96±30,23 mg/kg) do que na chuva (139,1±60,45 mg/kg). Em caprinos os teores de Zn no soro foram de 12,01±9,61 μmol/L na seca e 11,6±4,93 μmol/L na chuva e os do fígado foram de 134,2±26,58 mg/kg na seca e 131,45±76,55 mg/kg na chuva. Os teores séricos e hepáticos de Zn foram semelhantes em caprinos e ovinos. Considerando que os teores de Zn sérico encontram-se abaixo dos valores considerados como marginais e os hepáticos dentro do limite de normalidade, embora próximos ao limite inferior da referência, recomenda-se a suplementação com Zn, principalmente durante o período da seca.

Submitted by (edna.saturno@ufrpe.br) on 2016-10-14T12:17:11Z No. of bitstreams: 1 Leopoldo Mayer de Freitas Neto.pdf: 829417 bytes, checksum: 20a0e805e8a2d62a9b139566d0ef9786 (MD5) / Made available in DSpace on 2016-10-14T12:17:11Z (GMT). No. of bitstreams: 1 Leopoldo Mayer de Freitas Neto.pdf: 829417 bytes, checksum: 20a0e805e8a2d62a9b139566d0ef9786 (MD5) Previous issue date: 2010-02-15 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / In this work, divided into three experiments, the aim was to verify the possibility to diagnose the pregnancy of does and ewes by different pathway access and diagnose the fetuses sex originated by natural mating and transfer of fresh and cryopreserved embryos. The examinations were carried out using an ultrasound scanner equipped with a linear transducer (6.0 and 8.0 MHz) used by transrectal and transabdominal via and a microconvex endocavitary (5.0 and 7.5 MHz) transducer used by transvaginal via. In the first experiment was verified the viability of the ultrasound examination by transretal, transabdominal and transvaginal via to diagnose pregnancy in goats (n = 240) and ewes (n = 320) at days 30th, 45th, 60th and 75th. In does and ewes the ultrasound examination was faster (P < 0.05) on days 30th and 45th of pregnancy, however by transabdominal via was faster on day 60th and 75th. In both species the time of pregnancy diagnose was greater (P < 0.05) on day 30th than the others days while this time was smallest (P < 0.05) at day 75th than day 45th and 60th. Independent of the examination via and the day of pregnancy the average time to diagnose the pregnancy was shorter (P < 0.05) in does than in ewes. Independing of the specie and the day of pregnancy the average time to diagnose the pregnancy by transrectal via was shorter (P < 0.05) than the other vias and the transvaginal via was shorter (P < 0.05) than the transabdominal one. In the second experiment, in order to improve the sexing of Boer fetuses (n = 123) by transrectal ultrasonography, the aim was to identify the migration period of the genital tubercle and its later differentiation into genital structures in fetuses derived from natural mating (TI) and fetus from fresh (TII), frozen (TIII) and vitrified (TIV) embryo transfer collected 7 days after breeding. Migration of the genital tubercle occurred earlier (P < 0.05) in TI (45.21 ± 3.31 days) than in TII (48.50 ± 3.70 days), TIII (48.93 ± 3.61 days) and TIV (48.85 ± 3.23 days). The visualization of the scrotum, prepuce and vulva occurred earlier (P < 0.05) in TI (51.20 ± 2.56; 50.35 ± 1.59; 49.75 ± 1.73 days) than in TII (53.25 ± 2.02; 53.37 ± 1.92; 51.76 ± 2.10 days),TIII (53.37 ± 1.92; 52.31 ± 2.00; 51 ± 78 ± 2.22 days) and TIV (54.06±1.75; 52.46 ± 1.95; 51.91 ± 2.06 days). In the third experiment, in order to improve fetal sexing by ultrasonography in Dorper ewe breed (n = 130), the objective was to identifythe migration period of the genital tubercle and the period of the visualization of external genital structures in fetuses derived from natural mating (TI) and from fresh (TII), frozen (TIII) and vitrified (TIV) embryos transfer collected 7 days after breeding. Migration of the genital tubercle occurred earlier (P < 0.05) in TI (42.21±2.86 days) than in TII (43.98±3.00 days), TIII (44.97±1.83 days) and TIV (44.58±1.97days). The visualization of scrotal bag, prepuce and vulva occurred, respectively, earlier (P < 0.05) in TI (45.22±1.25; 45.95±1.53; 45.01±.103 days) than in TII (48.91±1.92; 48.52±1.41; 47.41±1.41 days), TIII (49.97±1.08; 49.18±2.00; 47.64±1.82 days) and TIV(50.12±1.66; 49.27±1.1.61; 47.93±1.92 days). The results allow to conclude that the pregnancy diagnose may be performed by transrectal, transabdominal and transvaginal via, as well as, that is faster by transrectal via and hat the time for diagnosing is shorterin advanced pregnancy and in goat specie. Is also possible to conclude, taking into consideration the final position of the genital tubercle, that goat fetal sexing can be done from the 55th day onward in fetuses produced by natural mating and from the 60th day onward in fetuses derived from cryopreserved embryos. It can also be concluded that, real-time ultrasonography is a reliable tool for fetal sex determination in sheep after Day 50 of pregnancy taking into account both the location of the genital tubercle and the identification of external genital structures. In ovine species the fetal sexing can be done from the 50th day onward in fetuses produced by natural mating and from the 55th day onward in fetuses derived from cryopreserved embryos. Finally is possible to conclude that real-time ultrasonography is a reliable tool for early pregnancy diagnose in small ruminant as well as that to identify the sex on the first 60 days of pregnancy. / Com este trabalho, dividido em três experimentos, objetivou-se verificar a possibilidade de realizar o diagnóstico de prenhez em cabras e ovelhas por diferentes vias de acesso e sexar fetos originados de monta natural e da transferência de embriões frescos e criopreservados. Os exames foram realizados com um aparelho de ultrassom equipado com um transdutor linear (6,0 e 8,0 MHz) utilizado pelas vias transretal e transabdominal e outro microconvexo (5,0 e 7,5 MHz) endocavitário utilizado por via transvaginal. No primeiro experimento verificou-se a viabilidade do exame ultrassonográfico pelas vias transretal, transabdominal e transvaginal para diagnosticar a gestação de cabras (n = 240) e ovelhas (n = 320) no 30o, 45o, 60o e 75o dia. Nas cabras e ovelhas, o exame ultrassonográfico pela via transretal foi mais rápido (P < 0, 05) no 30o e no 45o dia da gestação, mas, pela via transabdominal foi mais rápido no 60o e no 75o dia. Em ambas as espécies, a duração do diagnóstico de gestação foi maior (P < 0,05) no 30o dia do que nos demais, enquanto que a duração do diagnóstico no 75o dia foi menor (P < 0,05) do que no 45o e 60o. Independentemente da via de exame e do dia da gestação, o tempo médio para diagnosticar a gestação foi menor (P < 0,05) nas cabras do que nas ovelhas. Independentemente da espécie e do dia da gestação, o tempo médio para diagnosticar a gestação pela via transretal foi menor (P < 0,05) do que as demais e o da transvaginal foi menor (P < 0,05) do que o da via transabdominal. No segundo experimento, com a finalidade de aperfeiçoar a sexagem de fetos (n = 123) caprinos da raça Boer por via transretal, procurou-se identificar o período de migração do tubérculo genital e sua diferenciação nas estruturas da genitália de fetos derivados de monta natural (TI) e da transferência de embriões frescos (TII), congelados (TIII) e vitrificados (TIV) colhidos 7 dias após a cobertura. A migração do tubérculo genital ocorreu mais cedo (P < 0,05) no TI (42,21±2,86 dias) do que no TII (43,98±3,00 dias), TIII (44,97±1,83 dias) e no TIV (44,58±1,97 dias). A visibilização da bolsa escrotal, prepúcio e vulva ocorreu precocemente (P < 0,05) no TI (45,22±1,25; 45,95±1,53;45,01±1,03 dias) do que no TII (53,25 ± 2,02; 53,37 ± 1,92; 51,76 ± 2,10 dias), TIII (53,25 ± 2,02; 53,37 ± 1,92; 51.76 ± 2,10 dias) e TIV (54,06±1,75; 52,46 ± 1,95;51,91 ± 2,06 dias). No terceiro experimento, com o propósito de aperfeiçoar a sexagem de fetos (n = 130) ovinos da raça Dorper por via transretal, procurou-se identificar o período de migração do tubérculo genital e sua diferenciação nas estruturas da genitália de fetos derivados de monta natural (TI) e da transferência de embriões frescos (TII), congelados (TIII) e vitrificados (TIV) colhidos 7 dias após a cobertura. A migração do tubérculo genital ocorreu mais cedo (P < 0,05) no TI (45,21 ± 3,31 dias) do que no TII (48,50 ± 3,70 dias), TIII (48.50 ± 3.70 dias) e no TIV (48,85 ± 3,23 dias). A visibilização da bolsa escrotal, prepúcio e vulva ocorreu precocemente (P < 0,05) no TI (51,20 ± 2,56; 50,35 ± 1,59; 49,75 ± 1,73 dias) do que no TII (53,25 ± 2,02; 53,37 ± 1,92; 51,76 ± 2,10 dias), TIII (53,25 ± 2,02; 53,37 ± 1,92; 51.76 ± 2,10 dias) e TIV (54,06±1,75; 52,46 ± 1,95; 51,91 ± 2,06 dias). Os resultados permitem concluir que o diagnóstico de gestação pode ser realizado pelas vias transretal, transabdominal etransvaginal, bem como que é mais rápido pela via transretal, na gestação avançada e na espécie caprina. É também permissível concluir que, com base no posicionamento final do tubérculo genital, é recomendável sexar fetos caprinos provenientes de monta natural somente a partir do 55o dia de prenhez e a partir do 60o dia naqueles derivados da transferência de embriões frescos e criopreservados. Nos ovinos a sexagem fetal já pode ser efetuada a partir do 50o dia de prenhez nos fetos oriundos de monta natural e a partir do 55o dia naqueles originados da transferência de embriões frescos e criopreservados. Ainda é possível concluir que a ultrassonografia em tempo real é uma ferramenta importante para diagnosticar precocemente a prenhez nos pequenos ruminantes, assim como identificar o sexo fetal nos primeiros 60 dias de gestação

The selection on genetic values predicted from markers could substantially increase the rate of genetic gain in animals by increasing accuracy of prediction and reducing generation interval, especially for difficult to measure traits, such as feed efficiency. Feed efficiency is the most important trait in animal production due to its impacts on cost of production and environmental factors. Many metrics measure the feed efficiency, such as ratio of gain to feed (FER), the ratio of feed to gain (FCR) and residual feed intake (RFI). Nevertheless, in ovine, no study with the aim of understand the genetic variants or the accuracy of genomic estimated breeding value (GEBV) for feed efficiency traits was published yet. Moreover, before to apply the genomic information, it is necessary to understand and characterized the population structure, for instance, by linkage disequilibrium (LD). Both genome-wide association studies (GWAS) and genomic selection (GS) leverage LD between marker and causal mutation. Based on the above considerations, the aim of this study was to map LD in ovine, characterized by Brazilian Santa Inês sheep; to search genetic variants for feed efficiency traits (FER, FCR and RFI) through GWAS; and to verify the accuracy of GEBV for RFI. In total, 396 samples (animals) of Longissimus dorsi muscle were collect. A high-density panel of SNP (Illumina High-Density Ovine SNP BeadChip&reg;) comprising 54,241 SNPs was used to obtain the genotyping data. The phenotype data was comprised of 387 animals. The average LD between adjacent markers for two LD metrics, r² and |D\'|, were 0.166 and 0.617, respectively. The degree of LD estimated was lower than reported in other species and it was characterized by short haplotype blocks. Consequently, for genomic analyses, high-density panels of marker are recommended. Many markers were associated to feed efficiency traits in GWAS, mainly to RFI trait. Few candidate genes were reported in this study, highlighting NRF-1 (nuclear respiratory factor 1), which controls mitochondrial biosynthesis, the most important process responsible by a great fraction of the produced energy. Finally, we verified the accuracy of GEBV for RFI using few Bayesian regression models, and we found low accuracy, ranging from 0.033 (BayesB with &pi;=0.9912) to 0.036 (BayesA), which might be explained by the low relationship among animals and small training population. / A seleção com base nos valores genéticos genômicos preditos pode aumentar substancialmente a taxa de ganho genético em animais por meio do aumento da acurácia de predição e redução do intervalo de gerações, especialmente para características de difícil e/ou onerosa mensuração, como eficiência alimentar. A eficiência alimentar é uma das características mais importantes na produção animal devido principalmente aos seus impactos econômicos e ambientais. Muitas métricas representam a eficiência alimentar, por exemplo: a relação do ganho de peso e consumo alimentar (EA), a proporção do consumo alimentar e ganho de peso (CA) e o consumo alimentar residual (CAR). Em ovinos, nenhum estudo com o objetivo de buscar variantes genéticas ou verificar a acurácia do valor genético genômico estimado para eficiência alimentar foi publicado. Adicionalmente, antes de aplicar a informação genômica, é necessário compreender e caracterizar a estrutura da população, como por meio do desequilíbrio de ligação (LD). O estudo de associação genômica (GWAS) e seleção genômica (GS) consideram o LD entre marcador e a mutação causal. Com base nas considerações acima, o objetivo deste estudo foi mapear o LD em ovinos, caracterizado pela raça ovina Santa Inês; localizar variantes genéticas para as características de eficiência alimentar (EA, CA e CAR) utilizando a abordagem GWAS; e verificar a acurácia da estimação dos valores genéticos genômico para o CAR. No total, foram coletadas 396 amostras (animais) do músculo Longissimus dorsi, para posterior genotipagem utilizando o painel de alta densidade (Illumina High-Density Ovine SNP BeadChip&reg;), compreendendo 54.241 SNPs. O banco fenotípico é composto por 387 animais. O LD médio entre marcadores adjacentes para duas métricas de LD, r² e |D\'|, foram 0,166 e 0,617, respectivamente. O grau de LD estimado foi menor que o relatado em outras espécies e foi caracterizado por blocos de haplótipos curtos. Consequentemente, para as análises genômicas são recomendados painéis de marcadores de alta densidade. No GWAS, foram encontrados muitos marcadores associados aos fenótipos, em especial, à característica CAR. Alguns genes candidatos foram relatados neste estudo, destacando-se o NRF-1 (fator respiratório nuclear 1), que controla a biossíntese mitocondrial, o processo mais importante responsável por grande parte da produção de energia. Finalmente, verificamos a acurácia do valor genético genômico estimado para o CAR usando modelos de regressão Bayesiana, e encontramos baixos valores para acurácia (0,033 a 0,036) o que pode ser explicado pelo baixo grau de relacionamento entre os indivíduos e tamanho reduzido da população de treinamento.

As células-tronco mesenquimais (MSCs) são células estromais não-hematopoiéticas que possuem capacidade de diferenciação, sendo capazes, de diferenciar em diversos tecidos. As MSCs residem em vários tecidos vêm sido isoladas de diferentes tecidos, tais como cartilagem, tendão, tecido adiposo, do vaso e sangue umbilical, além de tecidos fetais . O isolamento e caracterização das populações mesenquimais no modelo ovino se faz importante, visto que ele é usado em ensaios pré-clínicos ortopédicos . Nesse estudo foram utilizados 5 amostras de sangue de cordão umbilical e 5 amostras de tecido adiposo peri-renal, provenientes de 10 ovinos fêmeas adultas. As coletas foram realizadas através de cirurgia para que o material coletado fosse o mais asséptico possível. Essas amostras foram submetidas a diferentes protocolos de isolamento, com a finalidade de se testar o mais eficiente. Somente um protocolo de cada tecido mostrou-se eficiente no isolamento da MSCs, porém nenhuma dessas amostras manteve-se viável após a primeira passagem. / Mesenchymal stem cells (MSCs) are non hematopoietic stromal cells that are able to differentiate through several tissues . MCSs home in several tissues and are being isolated from different tissues, such cartilage, tendon, adipose tissue, vessels and umbilical blood, and also from fetal tissues . The isolation and characterization of mesenchymal cells in sheep are important, because it is used in orthopedic pre-clinical trials . In this study were used 5 samples of umbilical blood and 5 samples of perirenal adipose tissue from 10 female sheep. All the samples were obtained through surgery, to harvest aseptic samples. These samples were tested in different protocols to evaluate the more efficient. Just one protocol from each source showed significant results in isolation, although none of the samples survived trough the first passage.

La paratuberculose est une maladie entérique incurable causée par la bactérie Mycobacterium avium subsp. paratuberculosis (MAP) endémique dans la plupart des régions du monde. Cette affection ciblant les ruminants est responsable d’importantes pertes économiques dans les productions animales et son contrôle est particulièrement difficile. L’ingestion de matières contaminées (fèces, lait et colostrum) est considérée comme la voie principale de transmission. La situation épidémiologique de la paratuberculose dans les troupeaux ovins du Québec demeure inconnue. Ce projet visait à évaluer la prévalence de MAP et à caractériser les facteurs de risque associés aux statuts positifs des ovins du Québec. Au total, 2942 brebis provenant de 70 fermes ovines, dont 10 productions laitières et 60 productions de boucherie, ont été échantillonnées dans le cadre de ce projet entre février 2020 et mars 2022. La prévalence réelle des moutons positifs dans la province a été estimée à 8.3% (IC 95% 4.7-14.1%) alors que la prévalence apparente de troupeaux positifs a été estimée à 29.9% (IC 95% 25.7-34.1%). L’analyse multivariée des facteurs de risque a permis d’identifier la présence de signes cliniques comme facteur de risque à un statut positif individuel (RC = 3.0; IC 95% 1.3-7.4) et l’utilisation d’un logiciel de régie pour les données de santé des animaux comme un facteur protecteur à un statut de troupeau positif (RC = 0.1; IC 95% 0.0-0.6). Les résultats de cette étude permettront l’exploration de mesures de contrôle de l’infection dans les troupeaux ovins de la province. / Paratuberculosis is an incurable enteric disease caused by Mycobacterium avium subsp. paratuberculosis (MAP), which is endemic in most parts of the world. This disease affecting ruminants is responsible for significant economic losses in livestock production and is particularly difficult to control. Ingestion of contaminated material (e.g., faeces, milk, colostrum) is considered the main route of transmission of MAP. The epidemiological situation of paratuberculosis in Quebec sheep flocks remains unknown. Hence, the purpose of this project was to evaluate the prevalence of MAP as well as to characterize the risk factors associated with positive status within Quebec sheep. A total of 2942 ewes from 70 sheep farms, including 10 dairy and 60 meat farms, were sampled between March 2020 and March 2022. The true prevalence of positive sheep in the province was estimated at 8.3% (IC 95% 4.7-14.1%) while the apparent prevalence of positive flocks was estimated at 29.9% (95% IC 25.7-34.1%). Multivariable analysis of risk factors identified the presence of clinical signs as risk factor for a positive ewe status (OR = 3.0; 95% CI 1.3-7.4) and the use of animal health data management software as a protective factor for positive flock status (OR = 0.1; 95% CI 0.0-0.6). The results of this study will enable the implementation of MAP-tailored infection control measures in the province's sheep flocks.

Le virus Schmallenberg (SBV) appartient au genre Orthobunyavirus, au sein de la famille des Bunyaviridae. Ce nouveau virus, découvert en 2011 au nord-ouest de l’Europe, affecte les ruminants domestiques. Il est responsable de signes cliniques discrets chez les adultes et de malformations congénitales chez les nouveau-nés. Ces travaux de thèse s’inscrivent dans les projets d’étude de la pathogenèse de l’infection à SBV et de l’épidémiologie de la maladie, dans le cadre d’un programme de recherche européen sur le virus. Ce manuscrit inclut de nouvelles données, telles les cinétiques de la virémie et de la séroconversion chez les ovins et caprins, après infection expérimentale par SBV. La possibilité d’infection par SBV par voie vaginale est démontrée expérimentalement chez la chèvre. Après infection expérimentale de chèvres gestantes entre 28 et 42 jours de gestation, une mortalité fœtale ou des lésions du système nerveux central des fœtus peuvent survenir. Enfin, la sensibilité de plusieurs espèces de ruminants sauvages et exotiques de parcs zoologiques vis-à-vis de SBV est démontrée pour la première fois. / Schmallenberg virus (SBV) belongs to the genus Orthobunyavirus in the family Bunyaviridae. This new virus was discovered in 2011 in Northwestern Europe in domestic ruminants. Infection by SBV is associated with mild clinical signs in adult and congenital malformations in the progeny. In the scope of the European research program on SBV in the pathogenesis and epidemiology areas, the works included in this thesis provide new data about SBV infection in livestock and wild and exotic ruminants. The kinetics of viremia and seroconversion after experimental SBV infection are described in sheep and goats. This manuscript includes evidence of SBV infection via vaginal route in goats. Experimental SBV infection in pregnant goats between 28 and 42 days of gestation can lead to death or central nervous system lesions in fetuses. Evidence of susceptibility to SBV in several species of wild and exotic ruminants kept in zoos is described for the first time.

<p>Вирус катаралне грознице оваца наноси велике штете сточарској производњи због чега су правовремено препознавање болести и брза дијагноза есенцијални предуслови за адекватну реакцију ветеринарске службе. Из тих разлога, предмет ове докторске дисертације су анализе патолошких и хистолошких промена код животиња, серолошких и молекуларних метода потребних за идентификацију и ближу карактеризацију узрочника што је од примарног значаја за ефективност предузетих мера у борби против катаралне грознице оваца.<br />Серолошки, током активног надзора, пргледано је 944 узорка крвног серума оваца и 953 узорка крвног серума говеда, а потом, током пасивног надзора, 114 узорака крвног серума говеда, 302 узорка крвног серума оваца и 22 узорка крвног серума коза. Коришћена је компетитивна имуноензимска ELISA метода, којом је доказано присуство специфичних антитела код говеда, оваца и коза. Из пуне крви серопозитивних јединки, ланчаном реакцијом полимеразе уз коришћење Pan-BTV rRT-PCR протокола заснованог на публикацији Toussiant и сар., (2007) је доказано присуство секвенце вирусне РНК. У случају позитивних резултата спроведена је и конвенционална PCR метода, употребом One step PCR кита, уз коришћење два пара прајмера за серотип BTV-4 чиме је доказано да је овај серотип узрочник обољења у Црној Гори. Секвенцирањем и филогенетском анализом изолата утврђено је да припада западном топотипу BTV-4 и да је сродан, готово идентичан са изолатима земаља из окружења.<br />Од 302 овце, којима је узета крв за серолошку анализу, уочене су каратеристичне макроскопске и микроскопске промене код свих угинулих од катаралне грознице оваца. Испитивањем је обухваћено 20 угинулих оваца које су претходно, током испитивања за живота, поред манифестне клиничке слике, имале позитивне резултате серолошких и молекуларних испитивања. Након угинућа, овце су обдуковане при чему су констатоване и сликане макроскопске промене те издвојена ткива за хистолошке анализе. Ткива су фиксирана у 10% пуферизованом неутралном формалину, а потом уклапљена у парафин. Парафински исечци дебљине 5 микрометара су бојени хематоксилин-еозин методом. На промењеним органима доминирају едем, хиперемија и крварења. Хистолошки се запажају интензивне хеморагичне инфилтрације свих промењених органа, а поред тога и местимични некротични процеси у срцу и језику као и периваскуларни едеми лимфних органа са израженом<br />лимфоцитном деплецијом у лимфним чворовима, слезини и тимусу.</p> / <p>Virus kataralne groznice ovaca nanosi velike štete stočarskoj proizvodnji zbog čega su pravovremeno prepoznavanje bolesti i brza dijagnoza esencijalni preduslovi za adekvatnu reakciju veterinarske službe. Iz tih razloga, predmet ove doktorske disertacije su analize patoloških i histoloških promena kod životinja, seroloških i molekularnih metoda potrebnih za identifikaciju i bližu karakterizaciju uzročnika što je od primarnog značaja za efektivnost preduzetih mera u borbi protiv kataralne groznice ovaca.<br />Serološki, tokom aktivnog nadzora, prgledano je 944 uzorka krvnog seruma ovaca i 953 uzorka krvnog seruma goveda, a potom, tokom pasivnog nadzora, 114 uzoraka krvnog seruma goveda, 302 uzorka krvnog seruma ovaca i 22 uzorka krvnog seruma koza. Korišćena je kompetitivna imunoenzimska ELISA metoda, kojom je dokazano prisustvo specifičnih antitela kod goveda, ovaca i koza. Iz pune krvi seropozitivnih jedinki, lančanom reakcijom polimeraze uz korišćenje Pan-BTV rRT-PCR protokola zasnovanog na publikaciji Toussiant i sar., (2007) je dokazano prisustvo sekvence virusne RNK. U slučaju pozitivnih rezultata sprovedena je i konvencionalna PCR metoda, upotrebom One step PCR kita, uz korišćenje dva para prajmera za serotip BTV-4 čime je dokazano da je ovaj serotip uzročnik oboljenja u Crnoj Gori. Sekvenciranjem i filogenetskom analizom izolata utvrđeno je da pripada zapadnom topotipu BTV-4 i da je srodan, gotovo identičan sa izolatima zemalja iz okruženja.<br />Od 302 ovce, kojima je uzeta krv za serološku analizu, uočene su karateristične makroskopske i mikroskopske promene kod svih uginulih od kataralne groznice ovaca. Ispitivanjem je obuhvaćeno 20 uginulih ovaca koje su prethodno, tokom ispitivanja za života, pored manifestne kliničke slike, imale pozitivne rezultate seroloških i molekularnih ispitivanja. Nakon uginuća, ovce su obdukovane pri čemu su konstatovane i slikane makroskopske promene te izdvojena tkiva za histološke analize. Tkiva su fiksirana u 10% puferizovanom neutralnom formalinu, a potom uklapljena u parafin. Parafinski isečci debljine 5 mikrometara su bojeni hematoksilin-eozin metodom. Na promenjenim organima dominiraju edem, hiperemija i krvarenja. Histološki se zapažaju intenzivne hemoragične infiltracije svih promenjenih organa, a pored toga i mestimični nekrotični procesi u srcu i jeziku kao i perivaskularni edemi limfnih organa sa izraženom<br />limfocitnom deplecijom u limfnim čvorovima, slezini i timusu.</p> / <p>The Bluetongue virus (BTV) incurs great damage to the production of livestock, due to which timely recognition of the resultant disease and rapid diagnosis are essential prerequisites for an adequate reaction by the veterinary service. For these reasons, the subject of this doctoral dissertation are the analyses of pathological and histological alterations in animals, the serological and molecular methods necessary for identifying and closely characterizing the causative agent which is of primary concern for the efficacy of the measures taken in the fight against bluetongue disease.<br />Serologically, during active surveillance, 944 samples of sheep blood serum as well as 953 samples of cattle blood serum were examined, followed by 114 samples of cattle, 302 samples of sheep and 22 samples of goat blood serum during passive surveillance. Competitive ELISA was utilized to detect the presence of specific antibodies in cattle, sheep and goats. The presence of viral RNA sequences was confirmed in whole blood samples of seropositive individuals using the Pan-BTV rRT-PCR protocol as described in Toussiant et al. (2007). In the case of positive results conventional PCR analysis was also performed, using the One Step PCR kit with two primer pairs specific to the BTV- serotype, which showed that this serotype is the cause of the disease in Montenegro. Sequencing and phylogenetic analysis of the isolate determined that it belongs to the western topotype of BTV-4 and that it is related and practically identical to the isolates from neighbouring countries.<br />This work describes the macroscopic and microscopic changes found in sheep which died due to affliction with bluetongue disease. The study investigated a sample of 20 sheep for which premortem analysis, apart from the manifested clinical features, had confirmed positive serologic and molecular test results. The animals were dissected postmortem during which macroscopic alterations were identified and imaged, followed by tissue isolation for histological analysis. Tissues were fixed in 10% neutral buffered formalin and embedded in paraffin. Sections of 5&mu;m thickness were cut and stained with haematoxylin/eosin. The predominant organ changes identified included edema, hyperaemia and bleeding. Histologically intensive hemorrhagic infiltrations of all affected organs were observed as well as sporadic necrotic processes in the heart and tongue accompanied by perivascular edemas of the lymphoid organs with pronounced lymphocyte depletion within the lymph nodes,<br />spleen and thymus.</p>

Venous disease will affect 1-3% of the western world at some point in their lives, yet there are few effective treatments for the venous system [1]. One such disease is chronic venous insufficiency (CVI), a painful and debilitating illness that affects the superficial and deep vein valves of the legs. When the valves become incompetent they allow reflux and subsequent pooling of blood. Current clinical therapies are only moderately; and therefore, the need for a better solution remains. Prosthetic venous valves were constructed from a novel hydrogel biomaterial patented by Georgia Tech. The valves had flexible cusps similar to normal, anatomic venous valves. The purpose of this work was to evaluate the thrombotic potential of the GT venous valve in an in vitro study and to design a percutaneous delivery system. In vitro thrombosis model provides an appropriate intermediate step between valve development and in vivo analysis, which is necessary to determine the biocompatibility of the prosthetic device. The flow system was modified from a one-pass, flow-through thrombosis assay using whole blood [2] to mimic pulsatile physiologic conditions. Cessation of flow indicated thrombotic obstruction. Histological analysis was performed using H and E staining and Carstairs stain (specific for platelets). A group of valves were lined with Dacron to confirm the thrombotic potential of the system. All Dacron valves were occluded by thrombus connecting the polymer fibers with adherent platelets. Whole blood perfused through the GT prosthetic valves exhibited no thrombosis or platelet adherence. All GT valves were patent and competent after blood perfusion. H and E staining revealed no thrombus deposition on the GT vein valves. A percutaneous delivery system was designed after evaluating the GT valves for their compressibility and plastic deformation over time. Appropriate stents, catheters and sheaths were selected. As designed, this system will be utilized in an ovine trial of the valve. Due to the low in vitro thrombotic potential and strong history of PVA as a medical implant material, positive trial results are expected. With successful animal and human trials this valve can provide a potential intervention for the 7 million people suffering from CVI.

Les ultrasons focalisés permettent d’effectuer des traitements thérapeutiques ciblés dans le corps humain. Dans le domaine des applications cardiovasculaires, ils permettent de détruire des caillots sanguins susceptibles de se former dans le système vasculaire. Dans ce cas, les mécanismes de thrombolyse sont largement liés à la cavitation ultrasonore, dont la dynamique complexe reste un obstacle à l’élaboration d’un dispositif thérapeutique. Dans le cadre de cette thèse, un système permettant le contrôle temporel de l’activité de cavitation en régime pulsé a été développé puis caractérisé. Ce dispositif utilise un transducteur focalisé et un hydrophone avec une boucle de rétroaction pour réguler l’activité de cavitation. Alors qu’en régime non régulé l’activité de cavitation a un caractère très aléatoire, le système de régulation mis au point permet d’atteindre un niveau de cavitation souhaité de manière très reproductible et avec une bonne stabilité temporelle. L’application de ce dispositif à la thrombolyse ultrasonore a été testée in vitro sur des caillots de sang humain. Au dispositif précédent a été ajouté un système permettant de déplacer le caillot sanguin au niveau du foyer, ainsi qu’un conduit permettant de compter le nombre de fragments libérés par la destruction du caillot. En comparaison des essais en régime non régulé, les essais en régime régulé ont montré une excellente efficacité thrombolytique et une très bonne reproductibilité, tout en diminuant les intensités acoustiques utilisées pour lyser les caillots sanguins. En parallèle des essais in vitro, une campagne de thrombolyse ultrasonore in vivo a été mise en place afin de réaliser des essais sur un modèle animal d’ischémie aiguë de membre inférieur. Un dispositif ultrasonore extracorporel in vivo guidé par échographie et monté sur un bras robotisé 6 axes a été développé. Un modèle ovin de thrombose artérielle a également été développé. Les tests ont permis de valider, d’une part, la faisabilité du modèle de caillot artériel et, d’autre part, le concept de thrombolyse extracorporelle purement ultrasonore basée sur la cavitation inertielle régulée / Focused ultrasound can be used for therapeutic applications in the human body. In cardiovascular applications, they can destroy blood clots formed in the vascular system. In this case, thrombolysis mechanisms are related to ultrasonic cavitation, but the complex dynamics remains an obstacle to the development of a therapeutic device. In this thesis, a system for the temporal control of the pulsed cavitation activity has been developed and characterized. This device uses a focused transducer and a hydrophone with a feedback loop for regulating the cavitation activity. While cavitation activity has a random behaviour in non-regulated conditions, the control system developed achieves a desired level of cavitation with very reproducibly and with good temporal stability. The application of this device to the ultrasound thrombolysis was tested in vitro on human blood clots. In the previous device was added a system for moving the blood clot at the focal point, and a tube for counting the number of fragments released by the destruction of the clot. In comparison to uncontrolled regime, tests showed an excellent thrombolytic efficacy and a very good reproducibility, with reduced acoustic intensities. In parallel to the in vitro tests, ultrasound thrombolysis was tested in vivo on an animal model of acute limb ischemia. An extracorporeal ultrasound device, guided by ultrasound and mounted on a robotic arm, has been developed for in vivo investigation. An ovine model of arterial thrombosis has also been developed. Tests were used to validate the feasibility of the model of arterial clots and to validate in vivo the concept of purely ultrasonic extracorporeal thrombolysis based on inertial cavitation regulation system